Cardiomyocyte precursors from human embryonic stem cells

• US 7,425,448 B2
• Filed: 07/12/2002
• Issued: 09/16/2008
• Est. Priority Date: 07/12/2001
• Status: Active Grant

• Current Assignee: Asterias Biotherapeutics Incorporated (Lineage Cell Therapeutics, Inc.)
• Original Assignee: Geron Corporation
• Inventors: Xu, Chunhui
• Primary Examiner(s): Ton; Thaian N
• Application Number:
  US10/193,884
• Publication Number:
  US 20030022367A1
• Time in Patent Office: 2,258 Days
• Field of Search: 435/325, 435/363, 435/366, 435/374, 435/377, 435/404, 435/405, 435/395, 435/401, 435/372
• US Class Current: 435/377
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 2500/30 Organic components metal chelators C12N2500/10; calcitonin C12N2500/14; transferrin C12N2500/24
• CPC Class Codes: C12N 2500/44 Thiols, e.g. mercaptoethanol
• CPC Class Codes: C12N 2500/62 DMSO
• CPC Class Codes: C12N 2501/06 Anti-neoplasic drugs, anti-retroviral drugs, e.g. azacytidine, cyclophosphamide
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2501/385 of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR]
• CPC Class Codes: C12N 2501/999 Small molecules not provided for elsewhere
• CPC Class Codes: C12N 2503/02 Drug screening
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0657 Cardiomyocytes; Heart cells

Abstract

This invention provides populations human cells of the cardiomyocyte lineage. The cells are obtained by causing cultures of pluripotent stem cells to differentiate in vitro, and then harvesting cells with certain phenotypic features. Differentiated cells bear cell surface and morphologic markers characteristic of cardiomyocytes, and a proportion of them undergo spontaneous periodic contraction. Highly enriched populations of cardiomyocytes and their replicating precursors can be obtained, suitable for use in a variety of applications, such as drug screening and therapy for cardiac disease.

Claims

1. A system for the production of human cardiomyocytes from human embryonic stem cells comprising:

a first isolated cell population comprising at least 0.56×

10⁶ cells differentiated in vitro from a portion of a population of human embryonic stem cells,

wherein the first cell population proliferates in culture and comprises at least 5% cardiomyocytes, identifiable by the criteria that they have spontaneous periodic contractile activity and express at least one of the following markers from an endogenous gene;

cardiac troponin I (cTnI), cardiac troponin T (cTnT), or atrial natriuretic factor (ANF); and

a second isolated cell population comprising said human embryonic stem cells,

wherein the second isolated cell population can produce more human cardiomyocytes by in vitro differentiation.

2. The system of claim 1, wherein the first isolated cell population comprises at least 20% cardiomyocytes.

3. The system of claim 2, wherein the first isolated cell population comprises at least 60% cardiomyocytes.

4. The system of any of claims 1-3, wherein the marker from an endogenous gene is cardiac troponin I (cTnI).

5. The system of any of claims 1-3, wherein the marker from an endogenous gene is cardiac troponin T (cTnT).

6. The system of any of claims 1-3, wherein the marker from an endogenous gene is atrial natriuretic factor (ANF).

7. The system of any of claims 1-3, wherein at least 18% of the cardiomyocytes express the marker Ki-67.

8. The system of any of claims 1-3, wherein the first isolated cell population comprises cells genetically altered to express telomerase reverse transcriptase.